Objective To examine the expression of metastasis-associated in colon cancer 1 (MACC1) gene in ovarian cancer cell lines and investigate its effect on biological behaviors of ovarian cancer cells.Methods The expression of MACC1 was examined by qRT-PCR and Western blot analysis in four ovarian cancer cell lines inculding OVCAR3,ES-2,SKOV3 and HO-8910.When the MACC1 was transfected to OVCAR3 cells,fluorogenic quantitative PCR was used to filter and identify MACC1 gene after the efficient silencing.Changes of adhesion in the cells were analyzed by an adhesion assay.Transwell migration and invasion assay and in vitro vascular mimicry assay were used to detect migration,invasion and angiogenesis of OVCAR3 cells in vitro.Results The expression of MACC1 gene was higher in OVCAR3 compared to other cell lines.qRT-PCR confirmed that the expression of MACC1 was silenced successfully after transient transfected MACC1-siRNA into OVCAR3 cells.After successful silencing the MACC1 expression,the adhesion ability was inhibited to some degree.In transwell migration assay,the numbers of cells in upper chamber passing through the membrane in transfected group were less than control groups (245.5 ±12.8,500.3±16.5 and 496.3±13.1 respectively),while in transwell invasion assay,the numbers of cells in upper chamber passing through the membrane in transfected group were less than the negative group and control group (185.3±14.1,405.7±9.1 and 416.3±11.5 respectively),both with markedly differences among the three groups.In tube formation assay,the distrubition of HUVECs was diffused with less junctions,and the average number of complete tubular structure was decreased in transfected group compared to the corresponding controls.Conclusion RNA interference inhibits the expression of MACC1 and effectively inhibits the metastasis and invasion abilities of ovarian cancer cells in vitro,and MACC1 is expected to become the target gene of ovarian cancer treatment.

Adult ; Female ; Humans ; Melanoma ; diagnosis ; pathology ; Neurocutaneous Syndromes ; diagnosis ; pathology

0.05).The adverse effect of treatment group was significantly less than control group(P

Objective To discuss the effects of implants of different thread face angles on the primary stability of immediately loaded implant using the 3-dimensional finite element analysis with the models of the immediately loaded implants. Methods Using the commercial code of Pro/E software, Hypermesh software, and ABAQUS software we created 3-dimensional finite element models. The micromotions of the finite element models with different screw face angles (V-shape, buttress, square-shape and inverse buttress) were computed with the ABAQUS software. Results Concerning different thread face angles, the micromotion of buttress implant was the minimum and the micromotion of inverse buttress implant was the maximum with vertically loading; the micromotion of inverse buttress implant was the minimum and the micromotion of buttress implant was the maximum with horizontal loading. Conclusions Different screw-types have great influence on vertical interfacial micromotions but little influence on horizontal interfacial micromotions. There are two angles which are formed by top /bottom edge and the implants. The larger are the angles, the smaller are the vertical interfacial micromotions, but the weaker of the strength. Thus in designing the screw-type implants, we should consider the angles of thread faces and the strength.

OBJECTIVEThe B7-H1/PD-1 co-signaling pathway has recently been found to play a pivotal role in the immune evasion of tumor cells from host immune system. The aim of this study was to examine the B7-H1 and PD-1 expression and TILs status in gastric cancer and to elucidate the clinical relevance of B7-H1 and PD-1 to the pathogenesis of gastric carcinoma.

METHODSImmunohistochemistry and ANAE histochemical staining were used to investigate the in situ expression of B7-H1 and PD-1 and TILs status in the gastric tissues. RT-PCR was used to explore B7-H1 and PD-1 expression at the transcriptional level. The B7-H1 expression at protein level was detected by Western blot.

RESULTSExpression of B7-H1 and PD-1 was found to be increased in gastric carcinoma, but absent in normal gastric tissue. B7-H1 expression in gastric carcinoma was inversely correlated with TILs infiltration. B7-H1 but not PD-1 expression in tumor tissue was significantly correlated with some clinicopathhological variables including depth of invasion, lymph node metastasis and distant metastasis.

CONCLUSIONB7-H1 and PD-1 expressions are increased in gastric carcinoma. This signaling pathway may inhibit antitumor immune responses in gastric carcinoma. B7-H1 expression plays a critical role in the pathogenesis of human gastric carcinoma,and might be a promising prognostic marker and therapeutic target in the treatment of this disease.

Adult ; Aged ; Antigens, CD ; genetics ; metabolism ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; B7-H1 Antigen ; CD4-Positive T-Lymphocytes ; immunology ; Female ; Humans ; Lymphatic Metastasis ; Lymphocyte Subsets ; immunology ; Lymphocytes, Tumor-Infiltrating ; immunology ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Staging ; Programmed Cell Death 1 Receptor ; RNA, Messenger ; metabolism ; Stomach Neoplasms ; genetics ; immunology ; pathology

BACKGROUNDEstrogen deficiency results in loss of bone mass. Phytoestrogens are plant-derived non-steroidal compounds with estrogen-like activity that bind to estrogen receptors. The main aim of this study was to investigate the effect of the phytoestrogen puerarin on adult mouse osteoblasts.

METHODSOsteoblast cells were harvested from 8-month old female imprinting control region (ICR) mice. The effects of puerarin stimulation on the proliferation, differentiation and maturation of osteoblasts were examined. The production of nitric oxide (NO) and the expression of bone morphogenetic protein-2 (BMP-2), SMAD4, mitogen-activated protein kinases (MAPK), core binding factor α1/ runt-related transcription factor 2 (Cbfa1/Runx2), osteoprotegerin (OPG), and receptor activator of NF-κB ligand (RANKL) genes were analyzed. The activation of signal pathways was further confirmed by specific pathway inhibitors.

RESULTSThe osteoblast viability reached its maximum at 10(-8) mol/L puerarin. At this concentration, puerarin increases the proliferation and matrix mineralization of osteoblasts and promotes NO synthesis. With 10(-8) mol/L puerarin treatment, BMP-2, SMAD4, Cbfa1/Runx2, and OPG gene expression were up-regulated, while the RANKL gene expression is down-regulated. Concurrent treatment involving the (bone morphogenetic protein) BMP antagonist Noggin or the NOS inhibitor L-NAME diminishes puerarin induced cell proliferation, Alkaline phosphatase (ALP) activity, NO production, as well as the BMP-2, SMAD4, Cbfa1/Runx2, OPG, and RANKL gene expression.

CONCLUSIONSIn this in vitro study, we demonstrate that puerarin is a bone anabolic agent that exerts its osteogenic effects through the induction of BMP-2 and NO synthesis, subsequently regulating Cbfa1/Runx2, OPG, and RANKL gene expression. This effect may contribute to its induction of osteoblast proliferation and differentiation, resulting in bone formation.

Animals ; Bone Morphogenetic Protein 2 ; genetics ; physiology ; Cell Differentiation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Female ; Isoflavones ; pharmacology ; MAP Kinase Signaling System ; physiology ; Mice ; Mice, Inbred ICR ; Nitric Oxide ; physiology ; Osteoblasts ; drug effects ; metabolism ; Osteogenesis ; drug effects ; Phytoestrogens ; pharmacology ; RANK Ligand ; genetics

OBJECTIVETo investigate the effects of mineral trioxide aggregate (MTA) and calcium hydroxide on the proliferation and differentiation capacity of pulp cells of primary teeth.

METHODSPulp cells were isolated from the retained primary teeth without apparent root resorption and cultured. The cells of 4 - 8 passages were used in the study. Cell proliferation was detected by MTT array, von Kossa staining employed to observe the formation of mineralized nodules and mRNA expression of alkaline phosphatase (ALP) and dentin sialophosphoprotein (DSPP) determined by real time PCR.

RESULTSMTA-treated cells proliferated significantly faster than the other two groups (F = 1835.065, P < 0.01), while calcium hydroxide-treated cells grew slower than the control significantly (F = 1792.301, P < 0.01). The formation of mineralized nodules was found in both MTA-treated and calcium hydroxide-treated pulp cells. The number of mineralized nodules showed no significant difference between the two groups (P > 0.05). Either ALP or DSPP mRNA expression showed significant difference among the three groups (F = 349.651, P < 0.01; F = 1653.001, P < 0.01). MTA increased mRNA expression of ALP and DSPP in pulp cells (P < 0.01), whereas calcium hydroxide down-regulated them (P < 0.01).

CONCLUSIONSMTA is more suitable than calcium hydroxide as pulp-capping agent in primary teeth.

Aluminum Compounds ; pharmacology ; Calcium Compounds ; pharmacology ; Calcium Hydroxide ; pharmacology ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dental Pulp ; cytology ; drug effects ; Drug Combinations ; Humans ; Oxides ; pharmacology ; Silicates ; pharmacology ; Tooth, Deciduous ; drug effects

OBJECTIVETo investigate the possible protective effect and mechanism of ginsenoside Rb1 against oxidative damage and renal interstitial fibrosis on rats with unilateral ureteral obstruction (UUO).

METHODSIn total, 80 male rats were randomly divided into 4 groups, 20 in each group: the sham operated group (SOR), UUO group, UUO with ginsenoside Rb1 treatment group (treated with intraperitoneal injection of 50 mg/ kg daily) and UUO with Losartan treatment group (as the positive control, treated with 20 mg/kg by gastrogavage per day). The rats were randomly sacrificed on day 3, 7 and 14 after surgery, respectively. The histopathologic changes of renal interstitial tissues were observed with Masson staining. The mRNA of transforming growth factor beta 1 (TGF-beta 1), collagen I and fibronectin were reversed transcribed and quantified by Real-time PCR. Enzyme-linked immunosorbent assay was used to quantitatively detect TGF-beta 1 and 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels. P47phox protein expression was assessed by immunohistochemistry and Western blot analysis.

RESULTSIn the UUO model, the obstructed kidney showed typical features of progressive renal tubulointerstitial fibrosis, and the levels of TGF-beta1, collagen I and fibronectin increased (P<0.05). As compared with the UUO group, ginsennoside Rb1 significantly inhibited the interstitial fibrosis including tubular injury and collagen deposition, and decreased the levels of TGF-beta1 (P<0.05). Ginsenoside Rb1 also inhibited the heme oxygenase (HO-1) and 8-OHdG, two markers of oxidative stress (P<0.05). Moreover, ginsenoside Rb1 suppressed the expression of p47phox, a subunit of nicotinamide adeninedinucleotide phosphate (NADPH) oxidase (P<0.05).

CONCLUSIONGinsenoside Rb1 can obviously inhibit renal interstitial fibrosis in rats with UUO, its mechanism possibly via against the oxidative damage and suppressing TGF-beta1 expression.

Animals ; Deoxyguanosine ; analogs & derivatives ; urine ; Drug Evaluation, Preclinical ; Fibrosis ; genetics ; metabolism ; prevention & control ; Gene Expression Regulation ; drug effects ; Ginsenosides ; therapeutic use ; Heme Oxygenase (Decyclizing) ; metabolism ; Kidney ; drug effects ; metabolism ; pathology ; Kidney Diseases ; etiology ; genetics ; pathology ; prevention & control ; Male ; Models, Biological ; NADPH Oxidases ; genetics ; metabolism ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Saponins ; therapeutic use ; Transforming Growth Factor beta1 ; genetics ; metabolism ; Ureteral Obstruction ; complications ; drug therapy ; genetics ; metabolism

Total saponins of Panax notoginseng (PNS) have been shown to ameliorate renal interstitial fibrosis. Ginsenoside Rg1, a panaxatriol saponin, is one of the major active molecules from PNS. The present study was undertaken to investigate the effect of ginsenoside Rg1 on renal fibrosis in rats with unilateral ureteral obstruction (UUO). The rats were randomly divided into 3 groups: sham-operation (n=15), UUO (n=15) and UUO with ginsenoside Rg1 treatment (n=15, 50 mg per kg body weight, intraperitoneally (i.p.) injected). The rats were sacrificed on Days 7 and 14 after the surgery. Histological examination demonstrated that ginsenoside Rg1 significantly inhibited interstitial fibrosis including tubular injury as well as collagen deposition. alpha-smooth muscle actin (alpha-SMA) and E-cadherin are two markers of tubular epithelial-myofibroblast transition (TEMT). Interestingly, ginsenoside Rg1 notably decreased alpha-SMA expression and simultaneously enhanced E-cadherin expression. The messenger RNA (mRNA) of transforming growth factor-beta1 (TGF-beta1), a key mediator to regulate TEMT, in the obstructed kidney increased dramatically, but was found to decrease significantly after administration of ginsenoside Rg1. Further study showed that ginsenoside Rg1 considerably decreased the levels of both active TGF-beta1 and phosphorylated Smad2 (pSmad2). Moreover, ginsenoside Rg1 substantially suppressed the expression of thrombospondin-1 (TSP-1), a cytokine which can promote the transcription of TGF-beta1 mRNA and the activation of latent TGF-beta1. These results suggest that ginsenoside Rg1 inhibits renal interstitial fibrosis in rats with UUO. The mechanism might be partly related to the blocking of TEMT via suppressing the expression of TSP-1.
Actins ; biosynthesis ; Animals ; Cadherins ; biosynthesis ; Collagen Type I ; genetics ; metabolism ; Fibronectins ; genetics ; metabolism ; Ginsenosides ; pharmacology ; Immunohistochemistry ; Male ; Nephritis, Interstitial ; drug therapy ; genetics ; metabolism ; pathology ; Panax notoginseng ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Smad2 Protein ; biosynthesis ; Thrombospondin 1 ; biosynthesis ; genetics ; Transforming Growth Factor beta1 ; biosynthesis ; genetics ; Ureteral Obstruction ; metabolism ; pathology

The research was conducted to study the breeding system of Prunella vulgaris L. Flowering dynamics was observed. Pollen viability, stigma receptivity, pollen-ovule ratio (P/O), out-crossing index (OCI) were measured. Bagging experiments were conducted. The results showed that the life span of one single flower was 1-2 days, the flowering span for the inflorescence of stalk was 7-14 days, the P/O was 1 046+/-148. 26, the OCI was 2. Combined with results of bagging experiment, the breeding system of P. vulgaris L. was mixed with cross-polination and self pollination. In the absence of pollination insects, the pollination and fertilization can be accomplished with high seed setting rate, and the seeds have a relatively high germination rate.
Breeding ; Pollen ; growth & development ; physiology ; Pollination ; Prunella ; growth & development ; physiology ; Tissue Survival